ap2a antibody  (Developmental Studies Hybridoma Bank)

Journal: Frontiers in Pharmacology

Article Title: Hyperglycemia-independent neonatal streptozotocin-induced retinopathy (NSIR) in rats

doi: 10.3389/fphar.2024.1395887

Figure Lengend Snippet: Intravitreal STZ injection on P1 inhibits the proliferation of neonatal rat retinal progenitors and delays their cell cycle exit and differentiation. (A) Horizontal P4 retinal sections of indicated locations and groups were stained for nuclear (DAPI, blue) and cell proliferation (Ki67, green). (B) Quantification of Ki67+ cells per mm 2 area of P4 and P8 retinas. (C) Horizontal P8 retinal sections of indicated locations and groups were stained for nuclear (DAPI, blue) and cell proliferation (Ki67, green). (D) Flow cytometry analysis of P4 retinal cells. (E) Whole-mount or horizontal sections of P8 retinas of indicated groups were stained for nuclear (DAPI, blue) and cell type markers, including Ganglion cells (Brn3, green, whole-mount), Cone (ARR3, green), Horizontal cells (OC2, green), Amacrine cells (Ap2a, green), Rod (Rho, green), Bipolar cells (Chx10, green) and Müller glia (Sox9, green). (F) Quantifying all seven retinal cell types (relative to the control group, %). Error bars represent SD of measurements from three animals or three retinas of three animals (n = 3), and asterisks indicate significant differences between control and STZ-treated groups (* p < 0.05, ** p < 0.01, one-way ANOVA followed by Bonferroni’s correction). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; Ivit, Intravitreal injection; PI, Propidium iodide. Scale bar: 50 μm.

Article Snippet: Slides were incubated with blocking solution (1% donkey serum and 0.1% Triton X-100 in PBS) for 1 h, and then with primary antibodies including Ap2a (Santa Cruz, SC-8975); Chx10 (Bremner lab, University of Toronto); Cleaved caspase-3 (Cell Signaling 9661); Cone arrestin or ARR3 (Millipore, AB15282); γ-H2ax (Millipore, 05–636); Onecut2 or OC2 (R&D system AF6294); Ki67 (BD science Pharmingen 550609); PH3 (Upstate 05–598); Rhodopsin (Santa Cruz SC-57433) and Sox9 (Millipore AB5535), overnight at 4°C.

Techniques: Injection, Staining, Flow Cytometry, Control

Journal: bioRxiv

Article Title: DNA-guided transcription factor cooperativity shapes face and limb mesenchyme

doi: 10.1101/2023.05.29.541540

Figure Lengend Snippet: A . Schematic of endogenous tagging TFs with the FKBP12 F36V degron and V5 epitope tag in human embryonic stem cells (hESC) followed by differentiation into cranial neural crest cells (hCNCC) and treatment with or without dTAG V -1. The ALX4 gene was instead knocked out by a frameshift mutation. B . Confirmation of TF tagging and depletion upon dTAG V -1 addition by Western blot for V5 epitope, with CTCF as a loading control. IB, immunoblot. C . Confirmation of ALX4 knockout in three independent clones by Western blot, with HSP90 as a loading control. D . Homeodomain TFs bind DNA at TWIST1-bound sites at variable occupancies. Heatmap shows promoter-distal binding sites for TWIST1 and/or AP2a. Assay indicates whether data shown is from ChIP, CUT&RUN (C&R), or ATAC, and whether an endogenous or V5 antibody was used. Rows are ranked by the sum of the homeodomain signals from undepleted cells. In the scale bar, units are reads per genome coverage, except for ATAC data, which is in signal per million reads. E . Homeodomain TFs bind the Coordinator motif. The top enriched motif (by AME) is shown for each TF, with p-values in parentheses.

Article Snippet: Antibodies used include TWIST1 (Abcam, ab50887), V5 (Abcam, ab15828), H3K27ac (Active Motif, 39133), Flag (Sigma-Aldrich, F1804), AP2a (Cell Signaling, 3215), AP2a (Novus Bio, NB100–74359).

Techniques: Mutagenesis, Western Blot, Control, Knock-Out, Clone Assay, Binding Assay

Journal: Cell Reports Methods

Article Title: Efficient and scalable generation of primordial germ cells in 2D culture using basement membrane extract overlay

doi: 10.1016/j.crmeth.2023.100488

Figure Lengend Snippet: BMEx overlay differentiation promotes differentiation to hPGCLCs alongside amniotic ectoderm- and mesoderm-like cells (A) Uniform manifold approximation and projection (UMAP) plots showing cluster identification (ID) (left) and time period in days (right) using single-cell transcriptomics of several hPSC lines undergoing differentiation with BMEx overlay. (B) Heatmap showing expression levels of the top 12 DEGs of each cluster. (C) Expression of signature genes of cell types of interest on the UMAP plot from (A). (D) UMAP showing integrated single-cell transcriptomics data from EB differentiation method (UCLA2 from Chen et al. ) and BMEx overlay method, highlighting the cells of EB differentiation method (left) and the BMEx overlay method (right). (E) Expression of signature genes of cell types of interest on the UMAP plot from (D). (F) Immunofluorescence for TFAP2A, TFAP2C, and GATA6 (top left); GATA3, SOX17, and TFAP2C (top right); and TFAP2A, GATA6, and PDGFRA (bottom) at day 5 with BMEx overlay. In the top panels, the dashed box is magnified (right), showing separated channels. The bottom panels depict a maximum intensity projection (MIP) and render image with a digital cross section showing separated channels (bottom part). Scale bars: 50 μm. (G) Whole-mount immunofluorescence for TFAP2A, GATA6, and PDGFRA in human WG9 amnion. In the top panels, a digital cross section (right) shows separated channels. The bottom panels depict a MIP and render image, also shown from the side (bottom). Scale bars: 50 μm. AELC, amniotic ectoderm-like cell; AMLC, amniotic mesoderm-like cell; iMeLCs, incipient mesoderm-like cells; PELC, primitive endoderm-like cell; Progen, progenitor cells. See also Figure S2 .

Article Snippet: Mouse anti-AP2a (TFAP2A) (1:200) , Santa Cruz Biotechnology , Cat# sc-12726; RRID: AB_667767.

Techniques: Single-cell Transcriptomics, Expressing, Immunofluorescence

Journal: Cell Reports Methods

Article Title: Efficient and scalable generation of primordial germ cells in 2D culture using basement membrane extract overlay

doi: 10.1016/j.crmeth.2023.100488

Figure Lengend Snippet: BMEx overlay potentiates BMP4 signaling and increases expression of critical PGC specification factors (A) Volcano plot showing DEGs between hPSCs at day 0 and 2-differentiated progenitors with BMEx overlay. (B) Immunofluorescence for TFAP2A, SOX17, and PRDM1 at days 2, 3, and 5 with BMEx overlay in line M54. TFAP2A is shown on top as a single channel. Dashed box is magnified (below), showing separate channels. Scale bars: 50 μm. (C) Immunofluorescence for pSMAD1/5/9, GATA3, and SOX17 at days 2, 3, and 5 with or without BMEx overlay in line M54. pSMAD1/5/9 is shown on top as a single channel. Scale bars: 50 μm. (D) Immunofluorescence for TFAP2A, EOMES, and SOX2 in line F99 at day 2 without (a) or with (b) BMEx overlay or with addition of 10 ng/mL activin A (C) or 10 μM SB431542 (D). Scale bars: 50 μm. (E) Violin plots depict the quantification of the images in (D) as the mean fluorescence intensity in arbitrary units (A.U.) of TFAP2A (left) and EOMES (middle) in DAPI segmented areas (normalized to 1) per cell at day 2. The correlation between these two values per cell per condition was visualized in a scatterplot (right). (F) Cartoon summarizing the hPGCLC differentiation progression in the BMEx overlay method as well as the perturbations tested (from D and E), with key analyzed markers depicted. See also Figure S4 .

Article Snippet: Mouse anti-AP2a (TFAP2A) (1:200) , Santa Cruz Biotechnology , Cat# sc-12726; RRID: AB_667767.

Techniques: Expressing, Immunofluorescence, Fluorescence

Journal: Cell Reports Methods

Article Title: Efficient and scalable generation of primordial germ cells in 2D culture using basement membrane extract overlay

doi: 10.1016/j.crmeth.2023.100488

Figure Lengend Snippet:

Article Snippet: Mouse anti-AP2a (TFAP2A) (1:200) , Santa Cruz Biotechnology , Cat# sc-12726; RRID: AB_667767.

Techniques: Recombinant, Staining, Membrane, Transfection, Bicinchoninic Acid Protein Assay, Software, Microscopy

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development.

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Figure 2. GRHL2 is required for AP2a activity at SE loci (A) Enrichment of various chromatin states in relation to GRHL2 ChIP-seq coordinates. Chromatin states were defined by ChromHMM using previously published histone mark ChIP-seq and ATAC-seq datasets in hESCs treated with RA/BMP4 for 7 days. (B) Expression of GRHL2-bound genes across integrated scRNA-seq identities. (C) Distribution of ATAC-seq signal (score based on number of reads per bin) relative to GRHL2 binding sites in WT and GRHL2 KO cells. (D) AP2a motif enrichment joint UMAP.

Article Snippet: Sheared chromatin was incubated overnight at 4 C with appropriate antibodies, followed by incubation on rotator with 30 mL of pre-washed agarose G beads (Invitrogen) for 4h at 4 C. AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 mg per 25 million cells andGRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 mg per 25 million cells.

Techniques: Activity Assay, ChIP-sequencing, Expressing, Binding Assay

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development.

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Figure 3. AP2a restricts GRHL2 binding to appropriate target genes (A) Distribution of GRHL2 ChIP-seq signal (score based on number of reads per bin) relative to GRHL2 or AP2a binding sites in WT and AP2a KO cells. (B) Distribution of ATAC-seq signal (score based on number of reads per bin) relative to the new GRHL2 binding sites in WT or AP2a KO cells. (C) Enrichment of various chromatin states in relation to AP2a KO GRHL2 ChIP-seq coordinates. Chromatin states were defined by ChromHMM using histone mark ChIP-seq and ATAC-seq datasets in hESCs treated with RA/BMP4 for 7 days. (D) Percentage of GRHL2 peaks found at or connected to promoters or AP2a binding sites in both WT and AP2a KO cells. p values were calculated with two- tailed Fisher’s exact tests. **** indicates a p value <0.0001.

Article Snippet: Sheared chromatin was incubated overnight at 4 C with appropriate antibodies, followed by incubation on rotator with 30 mL of pre-washed agarose G beads (Invitrogen) for 4h at 4 C. AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 mg per 25 million cells andGRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 mg per 25 million cells.

Techniques: Binding Assay, ChIP-sequencing, Two Tailed Test

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development.

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Figure 4. Cleft lip/palate genetic variants prioritized by integrating multiomic GRHL2/AP2a functional datasets (A) Pipeline overview, which connects single nucleotide polymorphisms (SNPs) in enhancers to distal genes via chromatin looping. (B) Flow chart illustrating how genes of interest were filtered using functional data. (C) Clinical significance of pipeline-identified output genetic variants. (D) Histogram of the minor allele frequency or (E) CADD scores of the genetic variants. Red dashed lines indicate standard cutoffs for minor allele frequency (MAF <0.02) or CADD score (CADD R15) for identifying disease-associated genetic variants. (F) Proportion of pipeline input or output genetic variants which were found directly from GWAS studies or are in linkage disequilibrium (LD). Significance was calculated with a Fisher’s exact test, * indicates a p value <0.05. (G) The chromosomal location and (H) functional categories of pipeline-identified genetic variants. (I) Number of pipeline-identified genetic variants falling within GRHL2 or AP2a binding sites, or AP2a-dependent ATAC sites in WT (black) or AP2a KO cells (white). p values were calculated with two-tailed Fisher’s exact tests, **** indicates p value <0.0001. (J) Overlap of genetic variants identified in our day 7 RA/BMP4 cells compared to published neural crest studies.39,40

Article Snippet: Sheared chromatin was incubated overnight at 4 C with appropriate antibodies, followed by incubation on rotator with 30 mL of pre-washed agarose G beads (Invitrogen) for 4h at 4 C. AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 mg per 25 million cells andGRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 mg per 25 million cells.

Techniques: Functional Assay, Binding Assay, Two Tailed Test

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development.

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Figure 5. Cleft lip/palate genetic variants alter GRHL2 binding and downstream gene expression (A) Table of SNPs identified from previous GWAS studies showing whether they fall within GRHL2 or AP2a binding sites. (B) Feature linkage between the ABCA4 promoter and accessible chromatin from WT and GRHL2 KO single cell multiome datasets. Arc height corresponds to the absolute value of each linkage. Red = negative correlation and Blue = positive correlation. (C) Chromatin accessibility at the ABCA4 locus in each cell type identified by scATAC. (D) AP2a and GRHL2 ChIP-seq signal in WT and AP2a KO cells at the ABCA4 locus. (E) Raw cohesion HiChIP signal at the ABCA4/ARHGAP29 locus in WT and AP2a KO cells. (F) Genetic variant preference at the SNP rs1211213 for AP2a or GRHL2 DNA binding as identified by ChIP-seq. Boxes represent median with interquartile ranges. (G) Expression of ABCA4 or (H) ARHGAP29 from bulk RNA-seq data. Error bars represent mean +/SD, n= 2 biological replicates. (I) UMAP of ABCA4 or (J) ARHGAP29 expression from integrated scRNA-seq. (K) Raw sequencing data illustrating the single nucleotide change at rs1211213 made with CRISPR genome editing. (L) qPCR comparing homozygous vs heterozygous rs1211213 alleles. Bars indicated mean with SD. p values were calculated with two-tailed Student’s ttest, n= 5 biological replicates run in triplicate. **** indicates p value <0.0001.

Article Snippet: Sheared chromatin was incubated overnight at 4 C with appropriate antibodies, followed by incubation on rotator with 30 mL of pre-washed agarose G beads (Invitrogen) for 4h at 4 C. AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 mg per 25 million cells andGRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 mg per 25 million cells.

Techniques: Binding Assay, Gene Expression, ChIP-sequencing, HiChIP, Variant Assay, Expressing, RNA Sequencing, Sequencing, CRISPR, Two Tailed Test

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: GRHL2 is required for AP2a activity at SE loci (A) Enrichment of various chromatin states in relation to GRHL2 ChIP-seq coordinates. Chromatin states were defined by ChromHMM using previously published histone mark ChIP-seq and ATAC-seq datasets in hESCs treated with RA/BMP4 for 7 days. (B) Expression of GRHL2-bound genes across integrated scRNA-seq identities. (C) Distribution of ATAC-seq signal (score based on number of reads per bin) relative to GRHL2 binding sites in WT and GRHL2 KO cells. (D) AP2a motif enrichment joint UMAP. (E) CRISPR strategy and immunoblot confirming AP2a KO hESCs. (F) Expression of GRHL2 and AP2a-dependent genes across integrated scRNA-seq identities. (G) Overlap of GRHL2 and AP2a-dependent and bound genes. Significance was calculated by Fisher exact test, ∗∗∗ indicates p value <0.0001. (H) Heatmap of read counts from WT, GRHL2 KO, and AP2a KO RNA-seq. (I) Expression of TFAP2A or GRHL2 in hESCs or WT, GRHL2 KO or AP2a KO hESCs treated with RA/BMP4. Error bars represent mean +/−SD, n= 2 biological replicates. (J) Immunoblot showing levels of AP2a protein in WT and GRHL2 KO cells treated with RA/BMP4. (K) Distribution of ATAC-seq signal (score based on number of reads per bin) relative to AP2a binding sites in WT and GRHL2 KO cells. Distribution of AP2a ChIP-seq signal (score based on number of reads per bin) relative to (L) all transcription star sites or (M) AP2a binding sites in WT and GRHL2 KO cells. (N) Representative bedgraphs of AP2a ChIP-seq in WT or GRHL2 KO cells.

Article Snippet: AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 μg per 25 million cells and GRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 μg per 25 million cells.

Techniques: Activity Assay, ChIP-sequencing, Expressing, Binding Assay, CRISPR, Western Blot, RNA Sequencing

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: AP2a restricts GRHL2 binding to appropriate target genes (A) Distribution of GRHL2 ChIP-seq signal (score based on number of reads per bin) relative to GRHL2 or AP2a binding sites in WT and AP2a KO cells. (B) Distribution of ATAC-seq signal (score based on number of reads per bin) relative to the new GRHL2 binding sites in WT or AP2a KO cells. (C) Enrichment of various chromatin states in relation to AP2a KO GRHL2 ChIP-seq coordinates. Chromatin states were defined by ChromHMM using histone mark ChIP-seq and ATAC-seq datasets in hESCs treated with RA/BMP4 for 7 days. (D) Percentage of GRHL2 peaks found at or connected to promoters or AP2a binding sites in both WT and AP2a KO cells. p values were calculated with two-tailed Fisher’s exact tests. ∗∗∗∗ indicates a p value <0.0001. (E) Number of chromatin contacts in WT and AP2a KO cells as measured by cohesin HiChIP. (F) Chromatin contact strength at ectopic GRHL2 binding sites in WT and AP2a KO cells. Boxes represent the median with interquartile range and error bars represent the minimum and maximum. (G) Empirical cumulative distribution function of the log 2 FoldChange in gene expression of promoters bound or looped to GRHL2 or AP2a binding sites (n = 6,564, purple) compared to all protein coding genes (n = 19,923, black) in WT vs. AP2a KO cells. p value <0.01 calculated by Student’s two-tailed test. (H) UMAP of integrated WT and AP2a KO cells colored by cell type. (I) Fold change in AP2a KO cell type proportions compared to WT, as measured by scRNA-seq. (J) Model of GRHL2 and AP2a regulation of gene expression. GRHL2 acts by blocking neural gene expression and promoting AP2a binding at surface ectoderm genes. In the absence of AP2a, GRHL2 binds unrestricted to inappropriate loci. (K) MA plot of differential expression analysis between WT and GRHL overexpression bulk RNA-seq. Transcript expression is either unchanged (gray), increased (blue, >2-fold), or decreased (red, <2-fold). p values for differentially expressed genes were calculated using DESEQ2 with a cutoff of 0.05. All differential genes and statistics can be found in Table S5 . (L) Overlap of GRHL2-regulated genes and genes altered by GRHL2 overexpression. p value was calculated with a Fisher exact test. (M) UMAP of integrated WT and GRHL2 over expression cells colored by cell type. (N) Fold change in GRHL2 overexpression cell type proportions compared to WT, as measured by scRNA-seq.

Article Snippet: AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 μg per 25 million cells and GRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 μg per 25 million cells.

Techniques: Binding Assay, ChIP-sequencing, Two Tailed Test, HiChIP, Gene Expression, Blocking Assay, Quantitative Proteomics, Over Expression, RNA Sequencing, Expressing

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Cleft lip/palate genetic variants prioritized by integrating multiomic GRHL2/AP2a functional datasets (A) Pipeline overview, which connects single nucleotide polymorphisms (SNPs) in enhancers to distal genes via chromatin looping. (B) Flow chart illustrating how genes of interest were filtered using functional data. (C) Clinical significance of pipeline-identified output genetic variants. (D) Histogram of the minor allele frequency or (E) CADD scores of the genetic variants. Red dashed lines indicate standard cutoffs for minor allele frequency (MAF <0.02) or CADD score (CADD ≥15) for identifying disease-associated genetic variants. (F) Proportion of pipeline input or output genetic variants which were found directly from GWAS studies or are in linkage disequilibrium (LD). Significance was calculated with a Fisher’s exact test, ∗ indicates a p value <0.05. (G) The chromosomal location and (H) functional categories of pipeline-identified genetic variants. (I) Number of pipeline-identified genetic variants falling within GRHL2 or AP2a binding sites, or AP2a-dependent ATAC sites in WT (black) or AP2a KO cells (white). p values were calculated with two-tailed Fisher’s exact tests, ∗∗∗∗ indicates p value <0.0001. (J) Overlap of genetic variants identified in our day 7 RA/BMP4 cells compared to published neural crest studies. ,

Article Snippet: AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 μg per 25 million cells and GRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 μg per 25 million cells.

Techniques: Functional Assay, Binding Assay, Two Tailed Test

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet: Cleft lip/palate genetic variants alter GRHL2 binding and downstream gene expression (A) Table of SNPs identified from previous GWAS studies showing whether they fall within GRHL2 or AP2a binding sites. (B) Feature linkage between the ABCA4 promoter and accessible chromatin from WT and GRHL2 KO single cell multiome datasets. Arc height corresponds to the absolute value of each linkage. Red = negative correlation and Blue = positive correlation. (C) Chromatin accessibility at the ABCA4 locus in each cell type identified by scATAC. (D) AP2a and GRHL2 ChIP-seq signal in WT and AP2a KO cells at the ABCA4 locus. (E) Raw cohesion HiChIP signal at the ABCA4 / ARHGAP29 locus in WT and AP2a KO cells. (F) Genetic variant preference at the SNP rs1211213 for AP2a or GRHL2 DNA binding as identified by ChIP-seq. Boxes represent median with interquartile ranges. (G) Expression of ABCA4 or (H) ARHGAP29 from bulk RNA-seq data. Error bars represent mean +/−SD, n= 2 biological replicates. (I) UMAP of ABCA4 or (J) ARHGAP29 expression from integrated scRNA-seq. (K) Raw sequencing data illustrating the single nucleotide change at rs1211213 made with CRISPR genome editing. (L) qPCR comparing homozygous vs heterozygous rs1211213 alleles. Bars indicated mean with SD. p values were calculated with two-tailed Student’s t test, n= 5 biological replicates run in triplicate. ∗∗∗∗ indicates p value <0.0001.

Article Snippet: AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 μg per 25 million cells and GRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 μg per 25 million cells.

Techniques: Binding Assay, Gene Expression, ChIP-sequencing, HiChIP, Variant Assay, Expressing, RNA Sequencing, Sequencing, CRISPR, Two Tailed Test

Journal: iScience

Article Title: GRHL2 and AP2a coordinate early surface ectoderm lineage commitment during development

doi: 10.1016/j.isci.2023.106125

Figure Lengend Snippet:

Article Snippet: AP2A ChIP antibody (Santa Cruz: sc-12726) used at 20 μg per 25 million cells and GRHL2 ChIP antibody (Sigma Prestige: HPA004820) used at 6 μg per 25 million cells.

Techniques: Virus, Recombinant, Blocking Assay, Plasmid Preparation, Magnetic Beads, Gene Expression, Purification, Cloning, dsDNA Assay, Sequencing, Software, Fluorescence